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RNAi 表型受注射品系的遗传背景影响。

RNAi phenotypes are influenced by the genetic background of the injected strain.

机构信息

Department of Developmental Biology, Blumenbach Institute of Zoology, Göttingen Center of Molecular Biology, Georg-August-University Göttingen, Göttingen, Germany.

出版信息

BMC Genomics. 2013 Jan 16;14:5. doi: 10.1186/1471-2164-14-5.

Abstract

BACKGROUND

RNA interference (RNAi) is a powerful tool to study gene function in organisms that are not amenable to classical forward genetics. Hence, together with the ease of comprehensively identifying genes by new generation sequencing, RNAi is expanding the scope of animal species and questions that can be addressed in terms of gene function. In the case of genetic mutants, the genetic background of the strains used is known to influence the phenotype while this has not been described for RNAi experiments.

RESULTS

Here we show in the red flour beetle Tribolium castaneum that RNAi against Tc-importin α1 leads to different phenotypes depending on the injected strain. We rule out off target effects and show that sequence divergence does not account for this difference. By quantitatively comparing phenotypes elicited by RNAi knockdown of four different genes we show that there is no general difference in RNAi sensitivity between these strains. Finally, we show that in case of Tc-importin α1 the difference depends on the maternal genotype.

CONCLUSIONS

These results show that in RNAi experiments strain specific differences have to be considered and that a proper documentation of the injected strain is required. This is especially important for the increasing number of emerging model organisms that are being functionally investigated using RNAi. In addition, our work shows that RNAi is suitable to systematically identify the differences in the gene regulatory networks present in populations of the same species, which will allow novel insights into the evolution of animal diversity.

摘要

背景

RNA 干扰 (RNAi) 是一种强大的工具,可用于研究不易进行经典正向遗传学研究的生物体中的基因功能。因此,随着通过新一代测序全面识别基因的便利性的提高,RNAi 正在扩大可用于研究基因功能的动物物种和问题的范围。在遗传突变体的情况下,已知用于的菌株的遗传背景会影响表型,而这一点尚未在 RNAi 实验中描述过。

结果

在这里,我们在赤拟谷盗 Tribolium castaneum 中表明,针对 Tc-importin α1 的 RNAi 会根据注射的菌株而导致不同的表型。我们排除了脱靶效应,并表明序列差异不能解释这种差异。通过定量比较四个不同基因的 RNAi 敲低引起的表型,我们表明这些菌株之间没有普遍的 RNAi 敏感性差异。最后,我们表明在 Tc-importin α1 的情况下,差异取决于母体基因型。

结论

这些结果表明,在 RNAi 实验中必须考虑菌株特异性差异,并且需要对注射的菌株进行适当的记录。对于越来越多的新兴模式生物,使用 RNAi 进行功能研究,这一点尤为重要。此外,我们的工作表明 RNAi 适合系统地鉴定同一物种群体中存在的基因调控网络的差异,这将为动物多样性的进化提供新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f3d/3574008/4568d065395a/1471-2164-14-5-1.jpg

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